Use of fluorescence-activated cell sorter to isolate mutant mammalian cells deficient in an internal protein, dihydrofolate reductase
- 1 January 1985
- journal article
- research article
- Published by Springer Nature in Somatic Cell and Molecular Genetics
- Vol. 11 (1), 71-77
- https://doi.org/10.1007/bf01534736
Abstract
Dihydrofolate reductase-deficient mutants of Chinese hamster ovary cells have been selected based on their inability to bind a fluorescent derivative of methotrexate, a substrate analog. Nonfluorescent mutant cells were isolated from mutagenized populations using a fluorescence-activated cell sorter. After multiple rounds of sorting plus regrowth, the mutant cell frequency was increased from an initial 10−5 to greater than 0.9. This use of the cell sorter to isolate mutants deficient in an internal protein should be applicable to any gene product that is able to bind a fluorescent ligand tightly. The method has the advantage of allowing the screening of large numbers of cells and of selecting for partially expressed phenotypes.Keywords
This publication has 20 references indexed in Scilit:
- Deletion of the diploid dihydrofolate reductase locus from cultured mammalian cellsCell, 1983
- Amplification and expression of sequences cotransfected with a modular dihydrofolate reductase complementary DNA geneJournal of Molecular Biology, 1982
- Selection of hybridomas and hybridoma variants using the fluorescence activated cell sorterJournal of Immunological Methods, 1982
- The selection of wild-type revertants from methotrexate permeability mutantsSomatic Cell and Molecular Genetics, 1978
- Folate-dependent enzymes in cultured Chinese hamster cells: Folylpolyglutamate synthetase and its absence in mutants auxotrophic for glycine + adenosine + thymidineArchives of Biochemistry and Biophysics, 1977
- Mutant alleles for hypoxanthine phosphoribosyltransferase: Codominant expression, complementation, and segregation in hybrid Chinese hamster cellsSomatic Cell and Molecular Genetics, 1976
- Isolation and partial characterization of three methotrexate-resistant phenotypes from Chinese hamster ovary cellsSomatic Cell and Molecular Genetics, 1976
- A replica plating method for CHO cells using nylon clothCytogenetic and Genome Research, 1975
- Isolation and biochemical characterization of folate deficient mutants of chinese hamster cellsCell, 1974
- Cloning of mouse myeloma cells and detection of rare variantsJournal of Cellular Physiology, 1972